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目的构建微小隐孢子虫CP23真核表达载体pcDNA3.0-23。方法提取微小隐孢子虫基因组DNA,PCR扩增CP23基因片段,然后克隆至TA载体,通过PCR、酶切及测序鉴定后,将CP23基因片段用限制性内切酶切下,克隆至真核表达载体pcDNA3.0,构建pcDNA 3.0-23重组质粒。通过PCR、酶切对重组质粒pcDNA3.0-23进行鉴定。结果扩增出约340 bp的微小隐孢子虫CP23基因片段,并成功构建微小隐孢子虫CP23真核表达载体pcDNA3.0-23。结论微小隐孢子虫CP23真核表达载体pcDNA3.0-23的构建为微小隐孢子虫的基因疫苗研制奠定了基础。
Objective To construct the eukaryotic expression vector pcDNA3.0-23 of Cryptosporidium parvum. Methods Genomic DNA of Cryptosporidium parvum was extracted and the CP23 gene fragment was amplified by PCR. The fragment was then cloned into the TA vector. After identification by PCR, restriction enzyme digestion and sequencing, the CP23 gene fragment was cut with restriction enzyme and cloned into eukaryotic expression vector Vector pcDNA3.0, construct pcDNA 3.0-23 recombinant plasmid. The recombinant plasmid pcDNA3.0-23 was identified by enzyme digestion. As a result, a CP 340 gene fragment of Cryptosporidium parvum of about 340 bp was amplified and the eukaryotic expression vector pcDNA3.0-23 of Cryptosporidium parvum CP23 was successfully constructed. Conclusion The construction of CP23 eukaryotic expression vector of Cryptosporidium parvum lays the foundation for the development of gene vaccine for Cryptosporidium parvum.